重组质粒pcDNA3.1(-)-P6-gBCTL-TBK-1的免疫效果评价

doi:10.6040/j.issn.1671-7554.0.2016.238

摘要/Abstract

摘要： 目的观察以TBK-1为分子佐剂的重组质粒pc(pcDNA3.1-)-P6-gBCTL-TBK-1的免疫效果。方法将构建的真核表达质粒pc-P6-gBCTL-TBK-1转染HeLa细胞瞬时表达, Western blotting验证融合蛋白的表达。将重组质粒肌内注射免疫接种BALB/c小鼠,分别于第0、2、4周免疫接种3次,第5周采集免疫小鼠的眼眶静脉血,第8周处死小鼠后无菌分离小鼠脾脏。ELISA法检测免疫小鼠血清的特异性抗HSV-2gD-P6抗体滴度、γ干扰素(IFN-γ)、白细胞介素-18(IL-18)、白细胞介素-4(IL-4)和白细胞介素-6(IL-6)的浓度,MTT法检测脾淋巴细胞的增殖率,乳酸脱氢酶(LDH)法检测细胞毒性T淋巴细胞(CTL)的活性。结果以TANK binding kinase-1(TBK-1)为分子佐剂的重组质粒可刺激机体产生特异性的抗HSV-2gD-P6抗体,诱导分泌较高水平的IFN-γ、IL-4和IL-18,并能促进小鼠脾淋巴细胞的增殖和增强CTL活性。结论构建的HSV-2DNA疫苗pc-P6-gBCTL-TBK-1具有一定的免疫原性,能诱导产生一定强度的细胞免疫和体液免疫,TBK-1具有良好的分子伴侣作用。

关键词: 单纯疱疹病毒2型, 小鼠, TANK结合激酶-1, DNA疫苗, 近交 BALB/c

Abstract: Objective To investigate the immune effects of herpes simplex virus type 2(HSV-2 )DNA vaccine of pc(pcDNA3.1-)-P6-gBCTL-TBK-1 in BALB/c mice. Methods The eukaryotic expression plasmid of pc-P6-gBCTL-TBK-1 was constructed and transfected into HeLa cells, and the expressed protein was verified by Western blotting. The BALB/c mice were immunized by the plasmids through intramuscular injection for three times, in week 0, 2 and 4. In week 5, the levels of interferon(IFN-γ), interleukin-18(IL-18), interleukin-4(IL-4), interleukin-6(IL-6)and specific HSV-2gD-P6 IgG antibody were measured by ELISA. In week 8, spleen cells of vaccinated mice were separated and the proliferation of spleen lymphocytes were detected by MTT. The activity of cytotoxic T lymphocyte(CTL)was detected by lactic dehydrogenase(LDH)method. Results After being inoculated by the plasmid of pc-P6-gBCTL-TBK-1, the mice were induced to generate higher level of specific antibody, including IFN-γ, IL-18 and IL-4; the CTL kill cells activity and proliferation capability of splenic lymphocytes were improved. Conclusion The recombinant plasmid of pc-P6-gBCTL-TBK-1 has immunogenicity, which can induce the humoral and cellular immune responses, and TBK-1 may have good function of molecular chaperone.

Key words: Herpes simplex virus type 2, DNA vaccine, TANK Binding Kinase-1, Mice, Inbred BALB C

中图分类号:

R373.11

焦凤萍,王玉,于爱莲,田兆菊,杨树林. 重组质粒pcDNA3.1(-)-P6-gBCTL-TBK-1的免疫效果评价[J]. 山东大学学报（医学版）, 2017, 55(3): 75-78.

JIAO Fengping, WANG Yu, YU Ailian, TIAN Zhaoju, YANG Shulin. Immunogenicity of the eukaryotic expression vector pcDNA3.1(-)-P6-gBCTL-TBK-1[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2017, 55(3): 75-78.

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